Determination of retinoid activity by an enzyme-linked immunosorbent assay. |
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| Authors: | S Michel A Courseaux C Miquel J M Bernardon R Schmidt B Shroot S M Thacher U Reichert |
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| Institution: | Centre International de Recherches Dermatologiques GALDERMA (CIRD GALDERMA), Sophia Antipolis, Valbonne, France. |
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| Abstract: | In normal human keratinocytes, retinoic acid suppresses the expression of the plasma membrane associated enzyme transglutaminase (TGm) at the pretranslational level. This finding led us to develop an enzyme-linked immunosorbent assay (ELISA) for the evaluation of the biological activity of retinoids, i.e., natural and synthetic derivatives of vitamin A. In this assay, keratinocytes are cultured in a 96-well cluster in the presence of different retinoid concentrations. The expression of TGm is then quantified, without any extraction or purification step, using a TGm-specific monoclonal antibody and a peroxidase-conjugated secondary antibody. The dose-response curves obtained show this ELISA to be a sensitive and reproducible assay to determine the potency of retinoids. |
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